Combating side-effects

ABSTRACT

The invention relates to the use of active ingredients, which increase the concentration of pyrimidine-based elements for nucleic acid biosynthesis in the body, in particular to the use of pyrimidine nucleosides and/or prodrugs produced therefrom, for reducing the side-effects of inhibitors of nucleic acid biosynthesis or their precursors, in particular by activating the biosynthesis of mitochondrial DNA (mtDNA). The invention also relates to the use of said active ingredients, in particular pyrimidine nucleosides and/or prodrugs for producing pharmaceutical preparations for reducing the aforementioned side-effects and to combinations or products for administering active ingredients of this type, in particular pyrimidine nucleosides and/or prodrugs produced therefrom, comprising inhibitors of nucleic acid biosynthesis or their precursors. The invention further relates to methods for treating the side-effects of inhibitors of nucleic acid biosynthesis or their precursors using the aforementioned active ingredients, in particular pyrimidine nucleosides and/or prodrugs produced therefrom, or the aforementioned combinations or products and to corresponding pharmaceutical preparations. Side-effects of HAART (Highly Active Anti-Retroviral Therapy) and side-effects of other anti-viral nucleoside analogous agents, which inhibit the mitochondrial γ-polymerases, can in particular be prophylactically and/or therapeutically treated in this manner.

[0001] The invention relates to the use of active agents that raise theconcentration of pyrimidine-base building blocks for the biosynthesis ofnucleic acids in the body, specifically of pyrimidine nucleosides and/orprodrugs of them, for the reduction of side effects of inhibitors of thebiosynthesis of nucleic acids or their preliminary stages, specificallythrough activation of biosynthesis of mitochondrial DNA (mtDNA), the useof these active agents, specifically of pyrimidine nucleosides and/orprodrugs, for the manufacture of pharmaceutical preparations for thereduction of the named side effects, combinations or products for theadministration of such agents, specifically, of pyrimidine nucleosidesand/or prodrugs of them, with inhibitors of the biosynthesis of nucleicacids or their preliminary stages, methods for the treatment of sideeffects of inhibitors of the biosynthesis of nucleic acids or theirpreliminary stages, using the named active agents, specificallypyrimidine nucleosides and/or prodrugs of them, or the namedcombinations or products, and corresponding pharmaceutical preparations.

BACKGROUND OF THE INVENTION

[0002] The term “HAART” (Highly Active Anti-Retroviral Therapy)summarizes a series of methods of treatment, in connection with whichanti-retroviral chemotherapeutic substances, specifically inhibitingsubstances of Reverse Transcriptase (RT) and inhibiting substances ofHIV protease are administered in combination to combat AIDS. Typicaltreatment programs employ two nucleoside-analogous RT inhibitors(NRTI's) and an HIV protease inhibitor or a non-nucleoside-analogous RTinhibitor. At first, above all after bone marrow suppression and theaccompanying negative effects of some inhibitors of the reversetranscriptase on the blood picture and the immune system have beenestablished as side effects, in part along with neuropathies andmyopathies (also of the heart muscle), further side effects becameevident with the new forms of therapy. One of them is the so-calledlipodystrophy. This term covers a series of symptoms and side effects.While high cholesterol and triglyceride levels are often found withprotease inhibitors, further side effects have been observed with theso-called nucleoside-analogous, reverse-transcriptase inhibitors(NRTI's). Particularly noticeable are the physical changes, such as thedisappearance of subcutaneous fatty tissue in the face, which can leadto shrunken cheeks and sunken eye sockets. The subcutaneous fatty tissuecan also be reduced in other parts of the body (e.g., the extremities).In part simultaneously with this, but also in isolation, an abnormalincrease in fatty tissue may occur (e.g., in the breasts of the womanand man, intra-abdominally, and in the neck and on the back of the neck.In particular, the latter symptoms are a burden for the patients, whosuffer under the fact that, as a result of the therapy, people can “seethe disease.” Possible consequences of the changes in fat are, amongother things, changes in the metabolism in other metabolic system areas,e.g., in the glycohomeostasis through resistance to insulin, and changesin the fat metabolism, and with that, e.g., the composition of the bloodlipids. All of these changes are comprised in the term lipodystrophy. Afurther effect, especially of the NRTI's, consists in damage to theliver, especially in the direction of an increase in fat (micro- ormacro-vesicular steatosis, steato-hepatitis, up to and including liverfailure). In addition, numerous other syndromes appear, such as changesin spermatogenesis (reduced sperm count, reduced sperm quality,specifically reduced sperm motility), a surge in cases of pancreatitisand a syndrome of osteopenia, which can be associated with an increasein the lactate level. Further, disorders of the kidney tubuli have beenobserved, e.g., reduced phosphate resorption. Finally, numerous HIVpatients taking NRTI in long-term anti-retroviral therapy manifestdiminished aerobic capacity on the basis of a diminished oxygen intake.All of the above-named changes seem to be long-term side effects of thedifferent active agents used in the context of HAART, in connection withwhich most of the above-named problems seem to be those of long-termtherapy with NRTI's.

[0003] Because HAART makes available very effective therapies, however,there is a great need for forms of treatment of this nature. For thisreason, options are urgently needed to limit, reduce, or eliminate thesenew side effects, specifically lipodystrophy, disorders ofspermatogenesis and the sperm function, the imbalance in the mineralsalt content of the bones (above and below referred to as osteopenia),the more frequent appearance of pancreatitis, diminished aerobiccapacity and/or disorders of the kidney functions, as well as myopathiesand/or weakening of the cells of the immune system. Up to this time,primarily adjustments in nutrition and prophylaxis against side effectswere used as therapy, in addition to the intake of vitamins and mineralsand the intake of unsaturated fatty acids, carnitine and the glutathionprecursor N-acetylcysteine, without a noticeable effect.

[0004] A further side effect of NRTI's is hyperlacticemia, which canlead to lactic acidosis, with symptomatic and in part life-threateningacute forms. Discontinuing the NRTI's only leads slowly to recovery, andthe use of cofactors or antioxidants such as ubiquinone, antioxidants,carnitine, riboflavin, and thiamin, has only a limited effect.

[0005] The treatment of all the side effects named has, all in all, beenrather unsatisfactory up to now—the temporary discontinuance of NRTI'sand/or other inhibitors of the biosynthesis of nucleic acids or itspreliminary stages, because of the danger of the development ofresistance and progression of HIV-associated immune weakness, is for themost part undesirable, as is a change in medication.

[0006] Patients infected by other viruses, e.g., the hepatitis B virus,the hepatitis C virus, the CMV virus, other herpes viruses (e.g., thevarizella-zoster virus, and herpes simplex viruses, the Epstein-Barrviruses, HHV type 6 and HHV type 8) and by the JC-virus, can also betreated with nucleoside analogues (e.g., with Acyclovir, Valacyclovir,Famaciclovir, Brivudin, Ribavirin, Ganciclovir, Tenofovir, Cidofovir andAdefovir) and potentially suffer the same side effects above, or thelong-term medication of the HIV-therapeutica above can be intensifiedthrough the simultaneous therapy with the nucleoside analogues directedagainst these viruses. Individual side effects in the use of theseantiviral substances can be significantly more evident than those withNRTI's, e.g., manifestation of the disorder of the kidney function knownas Fanconi syndrome.

[0007] For this reason, there is an urgent need to combat theabove-named undesirable effects of these medications, specificallylipodystrophy, changes in the sperm and/or osteopenia, in addition,pancreatitis, disorder of the kidney functions, diminished aerobicendurance, liver-damage and/or lactic acidosis, as well as to combatmyopathies and to reactivate existing immune cells.

[0008] In U.S. Pat. No. 5,968,914, the treatment of bone marrowsuppression induced by AZT and the accompanying anemia, as well asddC-induced peripheral neuropathy, ulcers of the mouth, reduced numberof thrombocytes, and the side effects on the muscles, peripheral nervoussystem, immune system, and gastro-intestinal tract caused by anti-viraltherapy through the administration of acylated non-methylated pyridinenucleoside, is described. WO 00/11952 describes, among other things, thetreatment of the side effects of cancer chemotherapy, such as peripheralneuropathies, kidney ailments and fatigue caused by the administrationof pyrimidine nucleoside precursors.

GENERAL DESCRIPTION OF THE INVENTION

[0009] It has been established that a large number of the side effectsof NRTI's and of the above-listed other nucleoside-analogous anti-viralmedications, specifically lipodystrophy, changes in the sperm and/orosteopenia, in addition, liver damage (specifically steato-hepatitis),hyperlacticemia/lactic acidosis, pancreatitis, disorders of kidneyfunction and/or diminished aerobic endurance and/or in addition,weakening of the existing cells of the immune system, can be effectivelytreated with active agents, preferably pyrimidine nucleosides and/or theprodrugs of them, that increase the concentration of pyrimidine buildingblocks for the synthesis of nucleic acids in the body.

[0010] Without intending to exclude other effective means and mechanismsby this statement, the effect can be plausibly explained in thefollowing way: NRTI's inhibit the gamma-polymerase required formitochondrial DNA-replication, and thereby the synthesis ofmitochondrial respiratory chain subunits. This inhibition is based, onthe one hand, on the competition of the NRTI's with the naturalmitochondrial nucleotides and, on the other hand, on the fact that theNRTI's serve as substrates for the polymerase, and because they lack a3-beta-hydroxy group, they bring about a break in the chain. Theresulting reduction of the quantity of mitochondrial DNA (mtDNA) thenleads to a reduced synthesis of these subunits of the mitochondrialrespiratory chain, which are coded by the mtDNA. This leads to thediminishing of the activity of the dihydro-orotate dehydrogenase(DHODH), an enzyme that is involved with the synthesis of pyrimidinenucleotides. The reason is that the activity of the DHODH is coupled tothe activity of the respiratory chain: the synthesis of orotate, apreliminary stage for pyrimidine nucleotides, is only possible ifelectrons of dihydro-orotate can be transferred to ubiquinone (coenzymeQ). This goes on continuously only if respiratory chains III and IV arefunctional. Orotate is then normally transformed into uridinemonophosphate, from which all of the other pyrimidines arebiosynthesized. For this reason, it can be assumed that the inhibitionof the function of the respiratory chain as a result of the presence ofNRTI's and/or other anti-viral nucleoside-analogous substances, whichtarget other viruses, leads to a reduction of the intramitrochondrialpyrimidine nucleoside and nucleotide. In consequence of this, the NRTIconcentration increases relative to that of the pyrimidine nucleotides;what results is an intensified reaction with the gamma-polymerase. Avicious circle arises, which leads by virtue of a further reduction ofthe mtDNA to even less pyrimidine nucleotide synthesis, etc. The effectof the administration of the active agents described above and below,according to the invention, specifically pyrimidine nucleosides and/orprodrugs of them, breaks this vicious circle by making available anexogenous source of these building blocks for mtDNA. In this way, itsuccessfully reduces the side effects of the nucleoside analogues andother inhibiting substances of the biosynthesis of mitochondrial nucleicacids, specifically mtDNA, or where relevant, their preliminary stages,and specifically combats the long-term effects described above.

[0011] The invention, therefore, is based on the use of active agentsthat increase the concentration of pyrimidine base building blocks forthe synthesis of nucleic acids in the body, specifically of pyrimidinenucleosides and/or their derivates, in order to eliminate theundesirable side effects of inhibitors of the biosynthesis of nucleicacids or their preliminary stages, specifically on the use of pyrimidinenucleosides and/or their derivates for (complete or partial)compensation for the depletion of mtDNA caused by these inhibitors,specifically by NRTI's and/or other anti-viral nucleoside analogues, andthe accompanying side effects.

DETAILED DESCRIPTION OF THE INVENTION

[0012] The invention involves (i) the use of active agents that increasethe concentration of pyrimidine base building blocks for thebiosynthesis of nucleic acids in the body, specifically pyrimidinenucleosides and/or prodrugs of them, for the reduction of side effectsof inhibitors of the biosynthesis of nucleic acids or their preliminarystages, specifically of NRTI's and/or of other anti-viral nucleosideanalogues, specifically the use of pyrimidine nucleosides and/or theirprodrugs for the activation of the biosynthesis of mitochondrial DNA(mtDNA), i.e., for the (complete or partial) compensation for depletionof mtDNA (which is specifically based on the inhibition of mitochondrialgamma-polymerase) and is caused by these inhibitors, specifically NRTI'sand/or other anti-viral nucleoside analogues, and of the accompanyingside effects, primarily lipodystrophy, changes in the sperm and/orosteopenia, in addition, liver damage (primarily micro- ormacro-vesicular steatosis, steato-hepatitis, up to and including liverfailure), hyperlacticemia/lactic acidosis, pancreatitis, disorder of thekidney function, diminished aerobic endurance and/or inhibition of theactivity of existing immune cells; (ii) the use of these active agents,specifically pyrimidine nucleosides and/or prodrugs, for the manufactureof pharmaceutical preparations for the reduction the named side effects,specifically through activation of the biosynthesis of mitochondrial DNA(mtDNA); (iii) combinations of preparations or products for theadministration of such agents, specifically pyrimidine nucleosidesand/or the prodrugs of them, with inhibitors of the biosynthesis ofnucleic acids or their preliminary stages; (iv) methods for thetreatment of side effects of inhibitors of the biosynthesis of nucleicacids or their preliminary stages, specifically through the activationof biosynthesis of mitochondrial DNA (mtDNA), using the named activeagents, specifically pyrimidine nucleosides and/or prodrugs of them, orthe named combinations or products; and (v) corresponding pharmaceuticalpreparations.

[0013] The general concepts and symbols above and below preferably havethe meanings listed below, provided that nothing is stated to thecontrary. Insofar as the general concepts and symbols named above andbelow are used, these can be replaced independently of one another bythe more specific definitions, which results in preferred embodiments ofthe invention.

[0014] The invention can be used prophylactically (i.e., for complete orat least partial avoidance of the named side effects or parallel to atreatment with inhibitors of biosynthesis of nucleic acids or theirpreliminary stages, for example HAART, and/or in the context of therapyfor other virus infections), and/or in case side effects have alreadyappeared, it can be used therapeutically (for example parallel to atherapy with the named inhibiting substances, or in treatment-freeperiods of time, e.g., (specifically in the case of treatment for lacticacidosis and/or hepato-toxicity) after the discontinuation of theinhibiting substances; all this is encompassed in the present invention.In a preferred embodiment of the invention, active agents according tothe present invention may also be used for treatment, i.e., preventionand therapy, of the named side effects (specifically, insofar as theyapply to the mitachondriopathies caused by the anti-viral therapy), inthe fetus and after birth, particularly in the first year of life (e.g.,in the case of newborns), in the context of the perinatal transmissionprophylaxis with inhibitors of the biosynthesis of nucleic acids ortheir preliminary stages.

[0015] Active agents that increase the concentration of pyrimidinebuilding blocks for the biosynthesis of nucleic acids in the body are,in the first place, compounds which, under physiological conditions,have the capacity to overcome an inhibition of the biosynthesis ofmitochondrial DNA (mtDNA) completely or at least partially (compensationand/or activation), if by treatment with inhibitors of biosynthesis ofnucleic acids or their preliminary stages, specifically NRTI's and/orother anti-viral nucleoside analogues, this synthesis can be completelyor partially prevented (i.e., specifically prophylactically before,prophylactically or therapeutically during, and/or therapeutically afteran appropriate corresponding anti-viral treatment), specifically by theinhibition of dihydro-orotate dehydrogenase (DHODH). Among the activeagents are specifically those compounds that have the capacity toincrease the concentration of orotic acid, ribonucleosides,deoxyribonucleosides, ribonucleotides and/or deoxyribonucleotides,primarily uridine and, in addition, cytidine, in the body, specificallyin the blood plasma, cells or other organelles such as mitochondria, asituation which, as described above, makes possible an activation orreactivation of the synthesis of mtDNA. Included here are specificallycompounds like uridine-phosphorylase inhibitors, inhibiting substancesof the uridine secretion, compounds that compete in connection withuridine renal transport mechanisms, or pyrimidine nucleosides and/orprodrugs of them. It is also possible to combine two or more of theactive agents named above and below, be it in a fixed combination, orstaggered over a period of time.

[0016] Compounds that are regarded as uridine phosphorylase inhibitorsare, specifically, compounds such as 5-benzylbarbiturate derivates,5-benzylbarbiturate, 5-benzyloxybenzylbarbiturate, 5-benzyloxybenzyl- or5-benzyloxybenzylacetyl-1-[(1-hydroxy-2-ethoxy)methyl]barbiturate,5-benzyloxybenzyl-1-[(1,3-dihydroxy-2-propoxy)methyl]barbiturate,5-benzyloxybenzyl-1-[(1-hydroxy-3-amino-2-propoxy)methyl]barbiturate,5-benzyloxybenzyl-1-[(2-(3-carboxypropionyloxy)ethoxy)methyl]-barbiturate,5-benzyl-1-[(1-hydroxy-2-ethoxy)methyl]barbiturate,5-methoxybenzylacetylbarbiturate,5-benzyl-1-[(1,3-dihydroxy-2-propoxy)methyl]barbiturate,5-benzyl-1-[(1-hydroxy-3-amino-2-propoxy)methyl]barbiturate,5-benzyl-1-[(2-(3-carboxypropionyloxy)ethoxy)methyl]barbiturate,5-methoxybenzylacetyl-acyclobarbiturate, 2,2′-anhydro-5-ethyluridine,and acyclouridin compounds like 5-benzyl-substituted acyclouridinederivates, for example, benzylacyclouridine,benzyloxy-benzyl-acyclouridine, aminomethyl-benzyl-acyclouridine,aminomethyl-benzyl-oxybenzyl-acyclouridine,hydroxymethyl-benzylacyclouridine orhydroxymethyl-benzyloxy-benzyl-acyclouridine.

[0017] Others are also known, e.g., in U.S. Pat. No. 5,567,689 and thereferences listed therein, particularly U.S. Pat. No. 4,613,604, U.S.Pat. No. 5,077,280 and/or U.S. Pat. No. 5,141,943.

[0018] Among inhibitors of uridine secretion are those compounds thatinhibit the transport of uridine from the cells, particularly the renalclearance of uridine. This class particularly includesN,N′-bis[3-(3,4,5-trimethoxybenzoyloxy)propyl]-homopiperazine (Dilazep)orN,N′-dimethyl)-N,N′-bis[3-(3′,4′,5′-trimethoxybenzoxy)propyl]-ethylendiamine(hexobendine), or other compounds listed in U.S. Pat. No. 5,567,689.

[0019] Compounds that compete with uridine in renal transport mechanismsare in particular L-uridine, L-2′,3′-dideoxyuridine undD-2′,3′-dideoxyuridine.

[0020] Especially preferred (possibly also over other mechanisms thanthose which work as those described) are pyrimidine nucleosides and/orprodrugs of them.

[0021] Pyrimidine nucleosides, are, in the first place, theribonucleosides or the deoxyribonucleosides of uracil or cytosine,specifically uridine and/or cytidine, in addition of thymine, such asthymidine.

[0022] Prodrugs are, in the first place, the metabolic preliminarystages of the named pyrimidine nucleosides from which the pyrimidinenucleosides are manufactured and/or released in the body. Thosecompounds that are preferred are those in which one or several of thehydroxy groups in the ribose radical are esterified, in each case withan acyl radical, and/or amino groups (as in cytidine or deoxycitidine),when present, are acylated with acyl radicals, or intermediates of thepyrimidine nucleotide biosythesis, which enter distally into thepyrimidine biosynthesis of the DHODH (such as orotic acid or its esters,for example, alkylester).

[0023] In addition, prodrugs can be selected from among triphenyluridine, triphenyl cytidine, uridine- or cytidine-5′-monophosphate orprodrugs from them (e.g., mono- or dialkylesters, acyloxyalkylesters,alkoxycarbonylmethylesters, substituted ethyl- and propylesters,amidomethylesters, benzylesters, phenylesters, phosphonoamidates,cyclophosphatesters, such as cytidine diphosphocholine or uridinediphosphoglucose); and oligo- or polynucleotides are used with U and/orC as base building blocks, like homo- or heterodimeres (e.g., U-P-U,U-P-C, C-P-U or C-P-C, where P stands for the binding bivalentphosphoric acid radical).

[0024] Wherever the active agents are mentioned that increase theconcentration of pyrimidine base building blocks for the synthesis ofnucleic acids in the body, specifically pyrimidine nucleosides or theirderivates, this refers, insofar as the named compounds containsaliferous groups, to the named compounds groups in free form and/or inthe form of salts. Salts of pyrimidine nucleosides or their prodrugs areparticularly pharmaceutically usable salts. If basic groups are present,like amino or imino, these can form acid addition salts, for examplewith inorganic acids like sulfuric acid or halogen hydrides or withorganic acids, for example, carboxylic acids like acetic acid, orsulfonic acids like methane sulfonic acid. If acid groups are present,like carboxy or sulfo, these can form salts with cations, for example,of metals like alkaline or earth alkaline metals, e.g., potassium orsodium, or with amino- or ammonia compounds, ammonium ions orlow-alkylamines. Inner salts, too, can be formed, if both acid and basicgroups are present.

[0025] Halogen refers specifically to fluorine, chlorine, bromine, oriodine.

[0026] The prefix “low” means that the radical in question haspreferably up to 7, specifically up to 4, carbon atoms.

[0027] “In addition” means “in a broader, specifically less preferredembodiment of the invention.”

[0028] “To increase the concentration in the body” means specifically anincrease of the concentration in the blood plasma, cells or theirorganelles, such as mitochondria, in the case of warm-blooded species,primarily humans.

[0029] Alkyl is in particular C₁-C₂₅-alkyl, preferably low-alkyl. Thenamed radicals, such as alkanoyl, alkenoyl or alkinoyl, can be presentin linear form, or in addition, in case the number of C-atoms allows it,in singly or multiply branched form.

[0030] Acyl is in particular the group of a carboxylic acid (especiallypreferred) that is bound by means of carbonyl to the binding oxygen (or,as in the case of cytidine, additionally to nitrogen) or of anunsubstituted or substituted amino acid (preferred); an acyl radical ofa semi-ester of carbon dioxide linked by means of its carbonyl group tothe binding oxygen or in addition nitrogen (preferred); or, in addition,an amino carbonyl group or an N-substituted amino carbonyl group. Acylhas preferably up to 25, primarily up to 20 carbon atoms, if nototherwise indicated.

[0031] The preferred acyl of a carboxylic acid is in particularunsubstituted or substituted C₁-C₂₅-alkanoyl, C₃-C₂₅-alkenoyl orC₃-C₂₅-alkinoyl, in particular low-alkanoyl, octanoyl, nonanoyl,decanoyl, undecanoyl, dodecanoyl or palmitoyl, or, in addition,substituted low-alkanoyloxy, in which situation the substitutecomponents are, for example, selected from one or more radicals,preferably one to three radicals, in particular of one radical, in everycase independently of one another, selected from among hydroxy,low-alkoxy, phenoxy, naphthoxy, low-alkanoyloxy, phenyl-low-alkanoyloxy,such as benzoyloxy or phenylacetyloxy, halogen, such as fluorine,chlorine, bromine or iodine, in particular fluorine or chlorine,carboxy, low-alkoxycarbonyl, phenyl-low-alkoxycarbonyl, such asbenzyloxycarbonyl, carbamoyl, low-alkylcarbamoyl,hydroxy-low-alkylcarbamoyl, di-low-alkylcarbamoyl,bis(hydroxy-low-alkyl)carbamoyl, cyano, oxo, C₃-C₈-cycloalkyl, such ascyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, C₆-C₁₂-bicycloalkyl,such as decahydronaphth-2-yl, C₉-C₁₄-tricycloalkyl, such as 1- or2-adamantyl, C₄-C₈-cycloalkenyl, such as 1-cyclohexenyl or1,4-cyclohexadienyl; heterocyclyl, which preferably indicates asaturated, partially saturated, or unsaturated simple ring that contains3 to 7, preferably 5 to 7, ring atoms, and that contains up to fourheteroatoms, selected from among nitrogen, sulfur, and/or oxygen,preferably 1 or 2 of the named heteroatoms; in which case the ring iseither present as such or can be as much as doubly, preferably singly,benz-anelated, cyclopenta-, cyclohexa- or cyclohepta-anelated; and whichcan be unsubstituted or substituted, in particular by low-alkyl,low-alkanoyl, hydroxy, low-alkoxy, phenyl-low-alkoxy, such as benzyloxy,hydroxy-low-alkyl, such as hydroxymethyl, halogen, cyano and/ortrifluormethyl, e.g., pyrrolyl, 2,5-dihydropyrrolyl, furyl, thienyl,tetrahydrofuryl, cyclohepta[bipyrrolyl, pyrrolidinyl, imidazolyl,imidazolidinyl, pyrazolinyl, pyrazolidinyl, triazolyl, such as 1,2,3-,1,2,4-or 1,3,4-triazolyl, tetrazolyl, such as 1- or 2-tetrazolyl,tetrahydro-oxazolyl, tetrahydro-isoxazolyl, tetrahydro-thiazolyl,tetrahydro-isothiazolyl, indolyl, isoindolyl, chinolyl, isochinolyl,benzimidazolyl, benzofuranyl, pyridyl, pyrimidinyl, piperidinyl,piperazine-1-yl, morpholino, thiomorpholino, S,S-dioxothiomorpholino,1,2-dihydro- or 1,2,3,4-tetrahydrochinolyl, or 1,2-dihydro- or1,2,3,4-tetrahydroisochinolyl, in which case the radicals mentioned areunsubstituted or are substituted as above, in particular by low-alkyl,e.g., in 4-low-alkyl-piperazine-1-yl, such as 4-methyl- or4-ethyl-piperazine-1-yl, by low-alkanoyl, e.g., in4-low-alkanoyl-piperazine-1-yl, such as 4-acetyl-piperazine-1-yl, or byhydroxy-low-alkyl, e.g., in 5-hydroxymethylfuran-2-ylcarbonyl, and aryl,preferably C₆-C₁₄-aryl, e.g., phenyl, naphthyl, such as 1- or2-naphthyl, or fluorenyl, such as fluoren-9-yl, in which case aryl isunsubstituted or is substituted for example singly or multiply,preferably multiply, by low-alkanoyl, for example by low-alkyl, e.g.,methyl, halogen-low-alkyl, such as trifluormethyl or chlorine or brominemethyl, halogen, e.g., fluorine or chlorine, hydroxy, low-alkoxy, suchas methoxy, low-alkanoyloxy, carboxy, low-alkyloxycarbonyl,phenyl-low-alkoxycarbonyl, carbamoyl, mono- or di-low-alkylcarbamoyl,mono- or dihydroxy-low-alkylcarbamoyl, such as heterocyclyl-low-alkyl,in which case heterocyclyl is defined as above as a substituent oflow-alkanoyl, in particular heterocyclylmethyl, in which heterocyclyl isbound by a ring nitrogen atom, e.g., piperidinomethyl,piperazine-1-ylmethyl, 4-low-alkyl-piperazine-1-ylmethyl, such as4-methyl- or 4-ethyl-piperazine-1-ylmethyl;4-low-alkanoyl-piperazine-1-ylmethyl; such as4-acetyl-piperazine-1-ylmethyl, morpholinomethyl or-thiomorpholinomethyl, cyano and/or nitro, in particular phenyl, whichis substituted by one of the radicals named in p-position, e.g.,low-alkanoyl, such as formyl, acetyl, propionyl, pivaloyl or heptanoyl,such as n-heptanoyl, hydroxy-low-alkanoyl, e.g., beta-hydroxypropionyl,low-alkoxy-low-alkanoyl, e.g., low-alkoxyacetyl or low-alkoxypropionyl,such as methoxyacetyl or beta-methoxypropionyl,low-alkanoyloxy-low-alkanoyl, e.g., low-alkanoyloxyacetyl orlow-alkanoyloxypropionyl, such as acetoxyacetyl orbeta-acetoxypropionyl, halogen-low-alkanoyl, e.g., alpha-halogenacetyl,such as alpha-chlorine-, alpha-bromine-, alpha-iodine-,alpha,alpha,alpha-trifluar or alpha,alpha,alpha-trichloracetyl, orhalogenpropionyl, such as beta-chlorine- or beta-bromine-propionyl,carboxy-low-alkanoyl, e.g., carboxyacetyl or 3-carboxypropionyl,low-alkoxycarbonyl-low-alkanoyl, e.g., low-alkoxycarbonylacetyl orlow-alkoxycarbonylpropionyl, such as methoxycarbonylacetyl,beta-methoxycarbonylpropionyl, ethoxycarbonylacetyl,beta-ethoxycarbonylpropionyl, tert-butoxycarbonylacetyl orbeta-tert-butoxycarbonylpropionyl, carbamoyl-low-alkanoyl, e.g.,carbamoylacetyl or beta-carbamoylpropionyl,low-alkylcarbamoyl-low-alkanoyl, di-low-alkylcarbamoyl-low-alkanoyl,hydroxy-carboxy-low-alkanoyl, hydroxy-low-alkoxycarbonyl-low-alkanoyl,dihydroxy-carboxy-low-alkanoyl,dihydroxy-low-alkoxycarbonyl-low-alkenoyl, heterocyclyl-low-alkanoyl,for example pyrrolylcarbonyl, such as 2- or 3-pyrrolylcarbonyloxy,furyl-carbonyl, e.g., 2-furylcarbonyl,5-hydroxymethyl-furan-2-ylcarbonyl, thienylcarbonyl, e.g.,2-thienylcarbonyl, imidazolocarbonyl, such as 4-imidazolylcarbonyl,imidazolylacetyl, such as 4-imidazolylacetyl, imidazolylpropionyl, suchas 3-(4-imidazolylpropionyl, pyridylcarbonyl, e.g., 2-, 3- or4-pyridylcarbonyl, indolylcarbonyl, e.g., 2-, 3- or 5-indolylcarbonyl,1-methyl-, 5-methyl-, 5-methoxy-, 5-benzyloxy-, 5-chlor- or4,5-dimethylindolyl-2-carbonyl, chinolyl-carbonyl, such aschinoline-2-ylcarbonyl, pyrrolidinylcarbonyl, such aspyrrolidinyl-3-carbonyl, piperidinylcarbonyl, e.g., 2-, 3- or4-piperidinylcarbonyl, morpholino-carbonyl, thiomorpholino-carbonyl,morpholinoacetyl, thiomorpholinoacetyl, or 4-low-alkyl-piperazinoacetyl,such as 4-methyl-piperazinoacetyl, low-alkenoyl, e.g., acryloyl,vinylacetyl, crotonoyl or 3- or 4-pentenoyl, low-alkinoyl, e.g.,propinoyl or 2- or 3-butinoyl, C₃-C₈-cycloalkylcarbonyl orC₃-C₈-cycloalkylacetyl, e.g., cyclopropyl-, cyclobutyl-, cyclopentyl- orcyclohexyl-carbonyl, cyclopropylacetyl, cyclopentylacetyl orcyclohexylacetyl, phenyl-low-alkanoyl, e.g., benzoyl, phenylacetyl or3-phenylpropionyl, in which case phenyl is unsubstituted or mono- ormultiply substituted by low-alkyl, e.g., methyl, halo-low-alkyl, such aschlorine or bromine methyl, halogen, e.g., fluorine or chlorine,hydroxy, low-alkoxy, e.g., methoxy, piperidinomethyl,piperazln-1-ylmethyl, 4-low-alkyl-piperazine-1-ylmethyl, such as4-methyl- or 4-ethyl-piperazine-1-ylmethyl,4-low-alkanoyl-piperazine-1-ylmethyl, such as4-acetyl-piperazine-1-ylmethyl, morpholino-low-alkyl, such asmorpholinomethyl, thiomorpholino-low-alkyl, such as methyl, cyano and/ornitro, e.g., 4-chlormethyl-, 4-brom-methyl-, 4-fluor-, 4-chlor-,4-methoxy-, 4-morpholinomethyl-, 4-thiomorpholinomethyl-, 4-cyano- or4-nitrobenzoyl, or low-alkylphenylacetyl, such as 4-methyiphenylacetyl.Especially preferred are the acyl radicals of unsubstituted carboxylicacids, as defined above, or the acyl radicals of carboxylic acidsphysiologically present, such as glykoloyl, lactoyl, enolpyruvoyl,liponoyl, pantothenoyl, acetoacetoyl, p-aminobenzoyl,beta-hydroxybutyroyl, creatinoyl or orotoyl.

[0032] Preferred acyl of an acyl radical of a semi-ester of carbondioxide connected to the binding oxygen by its carbonyl group is e.g.,unsubstituted or substituted hydrocarbyloxycarbonyl, preferably with 2bis 20 C-atoms, in particular unsubstituted or, in addition, substitutedlow-alkoxycarbonyl, e.g., methoxy-, ethoxy- or tert-low-alkoxycarbonyl,such as tert-butoxycarbonyl, 2-halogen-low-alkoxycarbonyl, e.g.,2-chlorine-, 2-bromine-, 2-iodine- or 2,2,2-trichlorethoxycarbonyl,aryl-low-alkoxycarbonyl, e.g., arylmethoxycarbonyl, in which case arylpreferably has 6 to 14 carbon atoms, is unsubstituted or is for examplesingly or multiply, preferably singly, substituted by low-alkyl, e.g.;methyl, halogen-low-alkyl, such as trifluormethyl or chlorine- orbromine-methyl, halogen, e.g., fluorine or chlorine, hydroxy,low-alkoxy, such as methoxy, low-alkanoyloxy, carboxy,low-alkyloxycarbonyl, phenyl-low-alkoxycarbonyl, carbamoyl, mono- ordi-low-alkylcarbamoyl, mono- or di-hydroxy-low-alkylcarbamoyl,heterocyclyl-low-alkyl, in which case heterocyclyl is defined as aboveas a substituent for low-alkanoyl, in particular heterocyclylmethyl, inwhich case heterocyclyl is bound by a ring nitrogen atom,piperidinomethyl, piperazine-1-ylmethyl,4-low-alkyl-piperazine-1-ylmethyl, such as 4-methyl- or4-ethylpiperazine-1-ylmethyl, 4-low-alkanoyl-piperazine-1-ylmethyl, suchas 4-acetyl-piperazine-1-ylmethyl, morpholinomethyl orthiomorpholinomethyl, cyano and/or nitro, and in particular phenyl, 1-or 2-naphthyl, fluorenyl or by low-alkyl, e.g., methyl or tert-butyl,low-alkoxy, e.g., methoxy, ethoxy or tert-butoxy, hydroxy, halogen,e.g., fluorine, chlorine or bromine, and/or nitro mono- ormultiply-substituted phenylist, e.g., phenyl-low-alkoxycarbonyl, such asbenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 4-nitrobenzyloxycarbonyl,diphenyl-low-alkoxycarbonyl, such as diphenylmethoxycarbonyl,di-(4-methoxyphenyl)-methoxycarbonyl trityloxycarbonyl orfluorenyl-low-alkoxycarbonyl, such as 9-fluorenylmethoxycarbonyl or inaddition heterocyclyl-low-alkoxycarbonyl, in which case heterocyclyl isdefined as above as a substituent of low-alkanoyl, e.g.,furan-2-ylmethoxycarbonyl or pyridine-2-, -3- or -4-ylmethoxycarbonyl.

[0033] A preferred N-substituted amino carbonyl group as acyl carries onits nitrogen 1 to 2 substituents, which are selected independently fromone another from substituted or unsubstituted low-alkyl, in which casethe substituents are selected from those named above for substitutedlow-alkanoyl and can be present in the number defined there, thosesubstituents preferably being selected from among hydroxy, low-alkoxy,low-alkanoyloxy, phenyl-low-alkanoyloxy, such as benzoyloxy orphenylacetyloxy, halogen, such as fluorine, chlorine, bromine or iodine,in particular fluorine or chlorine, carboxy, low-alkoxycarbonyl,phenyl-low-alkoxycarbonyl, such as benzyloxycarbonyl, cyano, oxo andphenyl or naphthyl, which are unsubstituted or for example singly ormultiply, preferably singly, substituted with low-alkyl, e.g., methyl,halogen-low-alkyl, such as trifluormethyl or chlorine- orbromine-methyl, halogen, e.g., fluorine or chlorine, hydroxy,low-alkoxy, such as methoxy, low-alkanoyloxy, carboxy,low-alkyloxycarbonyl, phenyl-low-alkoxycarbonyl, cyano and/or nitro, inparticular phenyl, which is substituted with one of the named radicalsin p-position; in particular from unsubstituted low-alkyl, such asmethyl or ethyl; and aryl, which preferably has 6 to 14 carbon atoms,and is unsubstituted or for example singly or multiply, preferablysingly, substituted by low-alkyl, e.g., methyl, halogen-low-alkyl, suchas chlorine- or bromine-methyl, halogen, e.g., fluorine or chlorine,hydroxy, low-alkoxy, such as methoxy, low-alkanoyloxy, carboxy,low-alkyloxycarbonyl, phenyl-low-alkoxycarbonyl, halo-low-alkyl, such astrifluormethyl, cyano and/or nitro, in which case the nitrogen of thecarbamoyl group no longer serves to bear an aryl radical. Thedefinitions falling under the various definitions of acyl groups of anN-substituted carbamic acid and the aminocarbonyloxy radical canpreferably be omitted.

[0034] A substituted or unsubstituted amino acid in acyl is formed,preferably by the amino acid radicals (aminoacyl): of an alpha-, beta-,gamma- or delta-amino acid bound by the carbonyl of its carboxy groupand an oxygen atom, in particular of a natural alpha-amino acid with theL-configuration, such as it normally occurs in proteins, or of an epimerof such an amino acid, i.e., with the unnatural D-configuration, or itsD,L-isomer mixture; of a homologue of such an amino acid, e.g., in whichcase the amino acid side chain is lengthened or shortened by one or twomethylene groups, in which case the amino group is present in beta-,gamma- or delta-position and/or in which case a methyl group is replacedby hydrogen; of a substituted aromatic amino acid, in which the aromaticradical has 6-14 carbon atoms, e.g., of a substituted phenylalanine orphenylglycine, in which case the phenyl-substituent can be low-alkyl,e.g., methyl, hydroxy, low-alkoxy, e.g., methoxy, low-alkanoyloxy, e.g.,acetoxy, amino, low-alkylamino, e.g., methylamino, di-low-alkylamino,e.g., dimethylamino, low-alkanoylamino, e.g., acetylamino orpivaloylamino, low-alkoxycarbonylamino, e.g., carbon atoms, e.g.,benzyloxycarbonylamino or 9-fluorenylmethoxycarbonylamino, halogen,e.g., fluorine, chlorine, bromine or iodine, carboxy and/or nitro,occurring singly or multiply; of a benz-anelated phenylalanine orphenylglycine, such as alpha-naphthylalanine, or of a hydratedphenylalanine or phenylglycine, such as cyclohexylalanine orcyclohexylglycine.

[0035] These amino acid radicals can be substituted in free amino orhydroxy functions, as described above for amino acid radicals R or Rg.Especially preferred is the radical of an amino acid bound by thecarbonyl of its carboxy group and an oxygen atom, selected from amongglycine, alanine, 2-amino butyric acid, 3-amino butyric acid, 4-aminobutyric acid, 3-amino valeric acid, 4-amino valeric acid, 5-aminovaleric acid, 3-amino caproic acid, 4-amino caproic acid or 5-aminocaproic acid, valine, norvaline, leucine, isoleucine, norleucine(alpha-amino caproic-acid), serine, homoserine(alpha-amino-gamma-hydroxy butyric acid), threonine, methionine,cysteine, proline, trans-3- and trans-4-hydroxyproline, phenylalanine,tyrosine, 4-amino phenylalanine, 4-chlorphenylalanine, 4carboxyphenylalanine, beta-phenylserine, phenylglycine,alpha-naphthylalanine, cyclohexylalanine, cyclohexylglycine, tryptophan,indolin-2-carboxylic acid, 1,2,3,4-tetrahydroisochinolin-3-carbon acid,asparagine acid, asparagine, amino malon acid, amino malonacid-monoamid, glutamine acid, glutamine, histidine, arginine, lysine,8-hydroxylysine, ornithine, 3-aminopropan acid, alpha,gamma-diaminobutyric acid and alpha,beta-diaminopropionic acid; especially preferredis the radical of an aliphatic amino acid, selected from among alanine,valine, norvaline, leucine, 3-aminopropion acid, 2-amino butyric acid,3-amino butyric acid, 4-amino butyric acid, 3-amino valeric acid,4-amino valeric acid, 5-amino valeric acid, 3-amino caproic acid,4-amino caproic acid or 5-amino caproic acid and isoleucine or of anamino acid selected from among glycine, asparagine, glutamine,methionine, lysine, histidine, proline, camitine and phenylalanine, inwhich case (except in instances in which there is no asymmetric carbonatom present, e.g., as with glycine) each of the named amino acids inthe D-, L- or (D,L)-, preferably in the L-Form can be present, and anamino group is present, unsubstituted or singly or doubly, or triply,N-alkyliert, e.g., by low-alkyl, such as methyl, n-propyl or n-butyl, bypyridyl-low-alkyl, such as 2-, 3- or 4-pyridylmethyl, and/or byphenyl-low-alkyl, such as benzyl, in which case the camitine radical ispreferred and/or N-acylated, e.g., by unsubstituted or substitutedlow-alkanoyl, as defined for low-alkanoyl, primarily by acetyl,propionyl or pivaloyl, aryl-low-alkanoyl, e.g., phenyl-low-alkanoyl,such as benzoyl or phenylacetyl, by low-alkoxycarbonyl, such astert-butoxycarbonyl, or by aryl-low-alkoxycarbonyl, e.g.,phenylnlederalkoxycarbonyl, such as benzyloxycarbonyl. Of the latterradicals, the preferred ones are acyl groups of an unsubstituted orsubstituted amino acid selected from among aminoacetyl (glycyl),N-low-alkylaminoacetyl, N,N-di-low-alkylaminoacetyl,N-low-alkyl-N-phenyl-low-alkylaminoacetyl,N-low-alkyl-N-low-alkoxycarbonylaminoacetyl andN-phenyl-low-alkoxycarbonyl-N-low-alkylaminoacetyl, e.g.,N-methylaminoacetyl, N,N-dimethylaminoacetyl,N-methyl-N-(n-butyl)aminoacetyl, N-methyl-N-benzylaminoacetyl,N-methyl-N-[(2-, 3- or 4-)pyridylmethyl-aminoacetyl, such asN-methyl-N-3-pyridylmethylaminoacetyl,N-methyl-N-tert-butoxycarbonylaminoacetyl,N-benzyloxycarbonyl-N-low-alkylaminoacetyl, prolyl, histidyl, glutamyl,asparagyl and carnitinoyl, in which case the amino acid radicals (exceptin cases in which there is no asymmetric carbon atom present, e.g., asin the case of gly) are present, preferably in the (L)-, in addition, inthe (D)- or (D,L)-form.

[0036] The usable prodrugs or pyrimidine nucleosides according to thepresent invention are specifically those of the formulas I (uridine orthe prodrugs of it) or II (cytidine or the prodrugs of it),

[0037] in which X stands for hydrogen or —OR₂, and R₁, R₂, R₃ and, whenpresent, R₄, are selected independently of one another, from hydrogenand acyl (preferably the acyl radicals described as preferred).

[0038] The usable pyrimidine nucleosides or prodrugs according to thepresent invention are known in the art, commercially available or can bemanufactured in accordance with known methods, e.g. according to oranalogous to those in U.S. Pat. No. 5,968,914, U.S. Pat. No. 5,583,117,U.S. Pat. No. 5,567,689, WO 00/11952, or EP 0 604 368. The citedreferences are herewith incorporated by reference, specificallyregarding the acyl groups and their introduction, and, insofar asdescribed, the prodrugs for pyrimidine nucleosides and theiradministration and formulation.

[0039] Inhibitors of the biosynthesis of nucleic acids (like DNA or RNA)or their preliminary stages (for example, nucleotides, deoxynucleotides,or in a given case the mono-, di- or tri-phosphates from them)specifically include:

[0040] inhibiting substances of ribonucleoside-diphosphate-reductase,primarily hydroxy urea (hydroxycarbamide; e.g., Litalir®, Bristol-MyersSquibb), Didox or Trimidox;

[0041] inhibitors of purine biosynthesis, such as2-morpholinoethyl-(E)-6-(1,3-dihydro-4-hydroxy-6-methoxy-7-methyl-3-oxoisobenzofuran-5-yl)-4-methyl-4-hexenoate(Mycophenolat.Mofetil; e.g.,CellCept®, Hoffmann-LaRoche);

[0042] inhibitors of dihydrofolate-reductase, e.g., Methotrexat,Pyrimethamin, Proguanil, Cycloguanil or Trimethoprim; or

[0043] preferably, inhibiting substances of mitochondrialgamma-polymerase, specifically inhibiting substances of reversetranscriptase (RT), for example of HIV, specificallynucleoside-analogous RT inhibitors (NRTI's), first and foremost AZT*(Zidovudin=3′-azido-3′-deoxythymidin; e.g., Retrovir®;Glaxo-SmithKline), Didanosin* (ddl, 2′,3′-Dideoxyinosin; e.g., Videx®;BMS), Zalzitabin* (ddC, Dideoxycytidin; e.g., HIVID Roche®), Lamivudin*(2′-Deoxy-3′-thiacytidin, 3TC; e.g., Epivir®; Glaxo-SmithKline),Stavudin* (d4T=2′,3′-didehydro-2′,3′-dideoxythymidine, e.g., Zerit®;BMS), Abacavir. (e.g., Ziagen®; Glaxo-SmithKline); or d4C(2′,3′-didehydro-2′,3′-dideoxycytidin), or in addition FTC(Emtricitabin, Covirazil=fluoridated 3TC; Triangle/Abbott); DAPD(guanosine analogues; Triangle/Abbott) as well as its de-aminatedmetabolite DXG; Fozivudin tidoxil (thioether-lipid-Zidovudin conjugate);ACH-126,443 (b-L-Fd4C, Achillion); Fosphazid (Phosphonavir, PZT);Tenofovir* (PMPA), Lodenosin (FddA, ddA), BCH-10618 (BioChemPharm),BCH-20652/dOTC (3TC-relative, BioChemPharm), BCH-13520, in which caseall those compounds marked-with asterisks (“*”) are preferred in therespective case;

[0044] or, in addition, other anti-viral nucleoside analogues(inhibitors of further viruses), e.g., nucleoside-analogous inhibitingsubstances of the hepatitis B virus, of the hepatitis C virus, of theCMV virus, of other herpes viruses, e.g., varizella-zoster virus, of theherpes simplex viruses, of the Epstein-Barr virus, of the HHV type 6,andthe HHV type 8) as well as of the JC virus (e.g., Acyclovir,Penciclovir, Sorivudin, Valacyclovir, Famciclovir) (e.g., Famvir;Glaxo-SmithKline), Brivudin (e.g., Helpin®, Berlin-Chemie), Entecavir(BMS-200475), Trifluridin (TFT; e.g., Thilo®), Idoxuridin (IDU; e.g.,Synmiol®), but first and foremost Fieluridin, Ribavirin (e.g., Rebetol®;Essex), Ganciclovir (e.g., Cymeven®), Cidofovir (e.g., Vistide®) undAdefovir Dipivoxil (e.g., Adefovir®g; Gilead);

[0045] or combinations of two or more of these inhibiting substances,and/or in addition, combinations of one of the above-named inhibitingsubstances with inhibitors of dihydro-orotic acid-dehydrogenase, e.g.,with Leflunomid (e.g., Arava®; Aventis; on the basis of the longhalf-value period, it is specifically possible to reactivate cells ofthe immune system several days after the administration of Leflunomidthrough an administration according to the present invention) orBrequinar.

[0046] The medications (=pharmaceutical preparations) according to theinvention can be administered to warm-blooded species (humans andanimals) nasally, rectally, orally, parenterally, intramuscularly, orintravenously, and contain a dosage of the pharmacological active agentalone, or together with a significant quantity of a pharmaceuticallyapplicable vehicle. The dosage of the active agent depends on thewarm-blooded species, body weight, age and the individual's condition,the individual pharmokinetic facts, the disease to be treated, as wellas the mode of application.

[0047] The dosage of the active agents that increase the concentrationof pyrimidine base building blocks for the biosynthesis of nucleic acidsin the body in warm-blooded species, specifically in humans, isparticularly in the range between 1 mg and 15 mg per m\2 of body surfaceper day; for uridine phosphorylase inhibitors, the concentration ispreferably in the range of 10 mg to 1 g per day, divided into one tothree doses; for inhibiting substances of the uridine secretion, thedosage is preferably in the range between 1 to 5 mg/kg of body weightone to three times a day; for compounds that compete with uridine inrenal transport mechanisms, the doses administered are preferably in therange of 5 to 50 mg/kg of body weight; administered one to three timesdaily.

[0048] The pyrimidine nucleosides and/or prodrugs of them are preferablyadministered in such doses that the blood plasma concentrations can beadjusted to lie between 10 and 500 μM, specifically between 20 and 300μM, primarily between 25 and 250 μM, e.g., 50-200 μM. The determinationof the blood plasma concentration takes place according to methods whichare known in the art, preferably through use of HPLC at specific pointsin time after the administration of the pyrimidine nucleosides or theprodrugs from them, e.g., 1, 2, 3, 4 and 5 hours after administration,for example as described in U.S. Pat. No. 5,583,117.

[0049] Pyrimidine nucleosides and/or prodrugs of them, specificallyuridine, cytidine, primarily mono-, di- or tri-acyluridin or mono-, di-,tri- or tetra-acylcytidin, are preferably administered daily inquantities from 10 to 500 mg/kg of body weight, specifically between 50and 300 mg/kg daily, for example, divided into two or three separatedoses, which are ingested in intervals of between 6 and 12 hours. Theacyl derivates are more easily absorbed when taken orally, and aretherefore preferred for oral administration.

[0050] The medications for use in accordance with the inventionencompass specifically one or several of the mitochondrial reactivators,specifically those named as preferred, together with standardpharmaceutical vehicles.

[0051] The active agents can be used, e.g., for the manufacture ofpharmaceutical preparations that contain an effective quantity of theactive agent, together or in a mixture with a significant quantity ofinorganic or organic, solid or liquid pharmaceutically usable vehicles.

[0052] The invention also relates to a pharmaceuticalcompound(preparation) that is suitable for administration towarm-blooded species, specifically humans, for the treatment orprevention of the side effects named above and below, specifically asregards HAART, primarily in connection with the administration ofNRTI's, or in connection with the treatment with other anti-viralnucleoside analogues, including a quantity of a mitochondrialreactivator sufficient to effect a reduction or elimination of the sideeffects, together with at least one pharmaceutically acceptable vehicle.

[0053] The invention further relates to a method for the therapeutic orprophylactic activation of the biosynthesis of mitochondrial DNA (mtDNA)before, during, or after the administration of inhibitors of thebiosynthesis of nucleic acids or their preliminary stages, specificallythose named as preferred, specifically also for the prevention of andtherapy for primarily the side effects named below, in the first placethose mitochondriopathies caused by the depletion of mtDNA in thecontext of perinatal transmission prophylaxis with inhibitors of thebiosynthesis of nucleic acids or derivates of them; and/or a method forthe therapeutic or prophylactic treatment of the side effects ofinhibitors of the biosynthesis of nucleic acids or their preliminarystages, specifically those named as preferred, which result from aninhibition of the orotate dehydrogenase and/or the biosynthesis ofmtDNA, in connection with which the side effects are manifest,specifically those of lipodystrophy, changes in the sperm and/orosteopenia, in addition, liver damage (primarily micro- or macrovesicular steatosis, steato-hepatitis up to and including liverfailure), hyperlacticemia/lactic acidosis, pancreatitis, disorder of thekidney function (including Fanconi syndrome) and/or diminished aerobicendurance, which arise in connection with the use of inhibitors of thebiosynthesis of nucleic acids or their preliminary stages, primarily inconnection with HAART, particularly by the administration of NRTI's, orin connection with therapy with other anti-viral nucleoside analogues,in connection with which a therapeutically or prophylactically effectivequantity of an active agent that increases the concentration ofpyrimidine building blocks for the synthesis of nucleic acids in thebody, specifically one described as preferred, or a pharmaceuticallyusable salt of it, in a dosage effective for the treatment of the namedside effect, specifically to a warm-blooded species, e.g., a human, whorequires the treatment because of one of the named side effects.

[0054] The pharmaceutical preparations contain from approximately 1% toapproximately about 96%, preferably from approximately 20% toapproximately 90%, of one or several active agents that increase theconcentration of pyrimidine base building blocks for the biosynthesis ofnucleic acids in the body (referred to below as an “active agent”).According to the present invention, pharmaceutical preparations can be,e.g., in single dose form, solutions of infusions ready for use,ampoules, vials, suppositories, dragées, tablets, or capsules.

[0055] The pharmaceutical preparations in the present invention areprepared in known ways, e.g., by conventional processes for solutions,freeze-drying, mixing, granulating, or the preparation of dragées.

[0056] For parenteral use, it is preferable to use capsules or solutionsof the active agent, also suspensions, particularly isotonic aqueoussolutions or suspensions, which can be prepared before use, e.g., in thecase of lyophilized preparations, which contain the active agent aloneor together with a vehicle, e.g., manna sugar. The pharmaceuticalpreparations can be sterilized and/or contain processing materials,e.g., preservatives, stabilizers, moisturizers and/or emulsifiers,solvents, salts for the regulation of osmotic pressure and/or buffers oracids, e.g., citric acid, and are manufactured in a well-known manner,e.g. by means of conventional solution or freezing processes. The namedsolutions or suspensions can contain substances that increase viscosity,such as sodium carboxymethyl cellulose, carboxymethyl cellulose,hydroxypropylmethyl cellulose, silica gel, dextran orpolyvinylpyrrolidon.

[0057] Suspensions in oil contain as oleagenous components thevegetable, synthetic, or semi-synthetic oils that are customary forinjection purposes, particularly fatty acid esters, which, as acidcomponents, contain a long-chain fatty acid with 8-22, specifically12-22 carbon atoms, e.g., lauric acid, palmitic acid, stearic acid orarachidonic acid, or corresponding unsaturated acids, e.g., oleic acidor linoleic acid, in a given case with the addition of antioxidants suchas vitamin E, beta-carotene, or 3,5-di-tert-butyl-4-hydroxytoluol. Thealcohol component of these fatty acid esters has a maximum of 6 carbonatoms and is a mono-or multihydric, e.g., mono-, di- or tri-hydric,alcohol, e.g., ethanol, isopropyl alcohol, but primarily glycol andglycerine. As fatty acid esters, therefore, the following examples canbe named: ethyl oleate, isopropyl myristate, isopropyl palmitate,polyoxyethylene glycerintrioleate, triglycerides of saturated fattyacids of a chain length C₈ to C₁₂, but especially vegetable oils likesesame seed oil.

[0058] The manufacture of preparations for injection takes place in thecustomary manner under sterile conditions, as does the filling intoampoules or vials and the sealing of the containers.

[0059] Pharmaceutical preparations for oral use can be obtained bycombining the active agent with stable vehicles, if desired, granulatinga prepared mixture and, if desired, processing the mixture into tablets,dragée kernels, or capsules after the addition of suitable processingmaterials. In this connection, synthetic vehicles can be included inthem, which release the active agents in regular doses or cause them tobe diffused.

[0060] Suitable vehicles include specifically fillers like sugar, e.g.,lactose, saccharose, manna sugar, or sorbitol, cellulose preparations,or calcium phosphate, in addition, binders like starch paste, using cornor potato starch, gelatins, tragacanth, methyl cellulose,hydroxypropylmethylcellulose, sodium carboxymethyl cellulose and/orpolyvinylpyrrolidon, and/or, if desired, dispersion agents like theabove-named starches, in addition, carboxyl methyl starch, cross-linkedpolyvinyl pyrrolidon, agar, agaric acid or its salt, such as sodiumalginate. Additives are first and foremost viscosity regulators andlubricants, e.g., silica gel, talcum, stearic acid or its salts, asmagnesium or calcium stearate, and/or polyethylene glycol. Dragéekernels are provided with suitable coatings, if necessary, which resistthe stomach acids.

[0061] Capsules are two-part capsules of gelatin, as well as soft,closed capsules of gelatin and a softener, such as glycerine orsorbitol. The two-part capsules can contain the active agent in the formof a granulate, e.g., with fillers like lactose, binders like starches,and/or lubricants like talcum or magnesium stearate, and possibly withstabilizers. In capsules, the active agent is preferably dissolved orsuspended in suitable oleaginous processing materials like fatty oils,paraffin oil or liquid polyethylene glycols, in which case stabilizersand/or antibacterial agents can likewise be added. Stabilizers, likeemulsifiers, surfactants, or tensides, binders like starch paste,tragacanth, or methylcellulose, hydroxypropylmethyl cellulose, sodiumcarboxymethyl cellulose, cyclodextrin and/or polyvinylpyrrolidon, and/orantibacterial agents can be added. Possible emulsifiers include oleicacid, non-ionic tensides of the fatty acid type-polyhydroxy alcoholester like sorbitol monolaurate, -oleate, -stearate or -palmitate,sorbitan tristearate or -trioleate, polyoxyethylene addition compoundsof fatty acid-polyhydroxy alcohol esters, like polyoxyethylene sorbitanmonolaurate, -oleate, - stearate, -palmitate, -tristearate or-trioleate, polyethylene glycol fatty acid esters like polyoxyethylstearate, polyoxyethylene glycol-(300 or 400)-stearate, polyethyleneglycol-2000-stearate, or ethylene oxide-propylene oxide-block polymers.

[0062] Coloring or pigments can be added to the capsule or dragéecoatings, e.g., for the purposes of identification or indication ofdifferent active-agent doses.

[0063] The administration can be parenteral as well as intravenous (forexample, by intramuscular, subcutaneous, or specifically intravenousinjection, by infusion or, specifically if toxic symptoms appear, likephlebitis of a peripheral vein, by central vein catheter), specificallyin the case of acute lactic acidosis or liver failure, otherwiseenteral, specifically oral.

[0064] The invention also relates to combinations of (A) active agentsthat increase the concentration of pyrimidine base building blocks forthe biosynthesis of nucleic acids in the body, with (B) one or severalother active agents, specifically with anti-viral active agents as thosein HAART, preferably NRTI's, or with other anti-viral nucleosideanalogues used for the treatment of infections with other viruses, inparticular the above-named. In particular, the invention includescombinations or products that contain the active agents named under (A)and one or several of the active agents named under (B), either in afixed combination or staggered over a period of time, or forsimultaneous use in a joint kit.

[0065] Products in which the therapeutica are available in a joint kitwith the preparations that contain the mitochrondrial reactivator arealso suitable for the treatment of side effects at intervals intreatment using the anti-viral therapy (interruption of HAART,specifically interruption of the therapy with NRTI's or with othernucleoside analogues).

[0066] In addition to the named inhibitors of the biosynthesis ofnucleic acids or their preliminary stages, further therapeutica,specifically for the treatment of AIDS, can be used simultaneously or ina staggered manner over time, such as entry inhibitors, CD4-bindinginhibitors, chemokin receptor antagonists, HIV-protease inhibitors,CXCR-4 antagonists, fusion inhibitors, non-nucleoside-analogous RTinhibitors (e.g., MKC-442 (Emivirin), SJ-3366 (Samjin Pharmaceuticals),TMC120 (Tibotec), UC-781 (Uniroyal), PNU-242721 (Pharmacia & Upjohn,Calanolide-A (SarawakMed), DPC-963,083 (DuPont), UIC-94-003, AG-1549(Capravirin; Agouron), GW420867X (Glaxo-Wellcome), DPC-961 (DuPont)),pyrophosphate analogues, integrase inhibitors, zinc finger inhibitors,interferons and tat/Rev inhibitors; as well as further therapeutica suchas antibiotics (for example for therapy for secondary infections in thecase of AIDS) or other chemotherapeutica. Corresponding pharmaceuticalpreparations can likewise be kits, in Which several active agents forsimultaneous or staggered administration are jointly present aspharmaceutical formulations or as fixed combinations.

PREFERRED EMBODIMENTS OF THE INVENTION

[0067] Preferred embodiments of the invention can be found in the use ofmore specific definitions for general expressions and symbols in theabove embodiments. Further preferred embodiments are found in thedependent claims.

[0068] Where the claims discuss uses, it is also possible, instead, touse the corresponding combinations or products for the administration ofactive agents which the concentration of pyrimidine base building blocksfor the biosynthesis of nucleic acids in the body increase, specificallypyrimidine nucleosides and/or prodrugs of them, with inhibitors of thebiosynthesis of nucleic acids or their preliminary stages, the methodsfor the treatment of side effects of inhibitors of the biosynthesis ofnucleic acids or their preliminary stages using the named active agents,specifically pyrimidine nucleosides and/or prodrugs or namedcombinations or products, or corresponding pharmaceutical preparations,which defines the preferred embodiments of the invention in each case.

EXAMPLES

[0069] The following examples serve the purpose of explaining theinvention, without limiting its scope.

Example 1 Compensation of the ddC-Caused Depletion of mtDNA in LiverCell Lines Through Uridine

[0070] Cell culture. The HepG2 cell line, a human hepatoma cell line, istaken from the American Type Culture Collection (ATCC No. HB-8065). Itis cultured at 37° C. in a 5% CO₂ atmosphere. The culture medium isDulbecco's Modified Eagle Medium (DMEM-containing 4.5 g/l glucose, 110mg/l pyruvate, 4 mg/l nicotinamide, 4 mg/l pyridoxal hydrochloride, 4mg/l thiamin and 0.4 mg/l riboflavin), supplemented with 10% fetalbovine serum (50 ml FCS, Mycoplex PAA Laboratories, Linz). On day 1,2.7×10⁶ cells are placed in 75 ml culture flasks (Falcon, BectonDickinson, USA). On days 5, 10, 15, 20 and 25, the cells aretrypsinizated and counted; 2.7×10⁶ cells are again spread out. Themedium is changed on each the days of the trypsin treatment and on thethird day after the repeated spreading.

[0071] Materials: Riboflavin (vitamin B₂), thiamin (vitamin B₁),ascorbic acid (vitamin C), uridine and ddC are purchased from Sigma(USA). The concentrations for the uridine are given in the followingtables; ddC is used in a concentration of 177 nM, which corresponds tothe equilibrium plasma levels of patients under HAART. A vitamincocktail consisting of 7.125 mg/l riboflavin, 7.125 mg/l thiamin and142.5 mg/l vitamin C is used. These pharmacological concentrationsexceed the daily requirement by a factor of about one hundred, and meetthe safe upper limits according to the recommendations of the EuropeanUnion for these micro-nutrients.

[0072] Determining the intracellular lipids: Intracellular lipiddroplets' are stained by means of Oil-Red-O (Sigma, USA). The cells arelaid out on cover glasses, incubated with Oil-Red-O (Sigma, USA) for 30minutes and counter-stained with Meyer's hematoxylin (8 min.), thenwashed with water and embedded in glycerin gelatin.

[0073] Lactic acid: 1.5 ml of the remainder of the culture is removed ineach case immediately before the trypsinization and the L-lactate in itis determined enzymatically with the lactate test-kit from RocheDiagnostics (order No.1822837) in an automatic analysis device(Roche/Hitachi 917) according to the recommendations of themanufacturer.

[0074] Quantification of mitochondrial DNA (mtDNA) by means of SouthernBlot: The content of mtDNA is determined by a well-known Southern-Blottechnique (C. T. Moraes et al., “Quantitative Defects of MitochondrialDNA”, in: S. DiMauro and D. C. Wallace, “Mitochondrial DNA in HumanPathology”, Raven Press, New York 1993, Chapter 8, pages 97-108).Genomic DNA is extracted from the fatty tissue and cut with therestriction endonuclease Pvull. Five μg of the cut material areseparated electrophoretically on a 0.8% agarose gel and transferred to anylon membrane. The mtDNA is tested with a 12.9 kBp mtDNA fragment(which comprises the nucleotide positions 3470 and 16379 of human mtDNA(S. Anderson et al., Nature 9, 457-65 (1981), and which is marked bymeans stochastically binding primers with digoxigenin). In order to havean internal standard, marked kernel DNA (mDNA), which hybridized the18S-ribosomal DNA gene that is present in a large number of copies, issimultaneously applied as a probe with a second digoxigenin-conjugatedprobe. The signals for mtDNA and nDNA are visualized, according to therecommendations of the manufacturer (Boehringer Mannheim, Germany), witha monoclonal antibody against digoxigenin, which is conjugate toalkaline phosphatase. The signal intensities are quantified by computerby means of standard software. The mtDNA content is expressed as themtDNA/nDNA (M/N) ratio.

[0075] Results: Apart from a short phase of hesitation after thetrypsinization, the HepG2-cells show logarithmic growth during theentire 25-day incubation period.

[0076] Uridine in a concentration of 200 μM normalizes the survival ofthe HepG2-cells in the presence of ddC, as can be seen from Table 1:TABLE 1 Number of cells (in 100,000s) on the day of the trypsinization(day 0 = 100%) Day 0 Day 5 Day 10 Day 15 Day 20 Day 25 HepG2-cellswithout NRTI, — 129.2 121.5 118.5 121.5 127.4 without uridine (SD =15.8) (SD = 12.3) (SD = 13.7) (SD = 7.9) (SD = 13.8) ddC 177 nM — 110.098.5 92.1103.0 66.2  38.4 ddC 177 nM + uridine 200 μM — 122.4 132.0114.4 120.2 130.6 ddC 177 nM + vitamins — 124.0 106.4 58.8 43.4 —

[0077] Uridine diminishes the reduction of the mtDNA; see Table 2. TABLE2 Content of mitochondrial DNA, expressed as an M/N-ratio in % of thecontrol (HepG2-cells without ddC and without uridine) Day 0 Day 5 Day 10Day 15 Day 20 Day 25 ddC 177 nM 100.0 25.6 25.1 11.5 8.3 8.4 ddC 177nM + 100.0 74.7 60.4 64.2 53.1 67 uridine 200 M ddC 177 nM + 100.0 16.08.9 9.7 9.3 — vitamins

[0078] The production of lactic acid, which appears as the result ofincreased compensatory glycolysis and an NADH/NAD equilibrium shifted infavor of NADH because of the inhibition of the mitochondrial oxidativephosphorylation by the inhibitors of the gamma-polymerase, is diminishedby the uridine; see Table 3. TABLE 3 Lactate values in the medium inpercent of the control (without ddC and without uridine) - Day 0 = 100%Day 0 Day 6 Day 10 Day 15 Day 20 Day 25 ddC 177 nM — 109.2 129.5 220.1374.9 392.5 ddC 177 nM + — 101.5 99.1 100.7 106.6 — uridine 200 μM ddC177 nM + — 196.1 143.0 249.0 302.1 — vitamins

[0079] In measuring the intracellular lipids, on Day 20 there is a cleardiminution of steatotic changes in the presence of uridine. While in thepresence of 177 μM of ddC and 200 μM of uridine, the microscopic imageresembles that of the control without ddC and without uridine (manycells, little red color from Oil-Red-O), in the presence of 177 μM ddC(without uridine) clearly fewer cells are found, which however, arerelatively large and red-colored (=containing fat). In the presence of177 μM ddC and the vitamin cocktail, the image attained resembles thatin the presence of 177 μM ddC without uridine. Thus it is solely thepresence of the uridine which can compensate for the side effects ofddC. Supplementing with vitamins, on the other hand, produces norecognizable favorable effect (see also Tables 1 to 3).

Example 2 Compensation for the NRTI-Stipulated Depletion in mtDNA inLiver Cell Lines Through Uridine

[0080] Analogously to Example 1, experiments were carried out with ddIand with d4T instead of ddC.

Example 3 Example of Formulation for an Infusion Solution with Uridine

[0081] In a concentration of 10 mg/ml, uridine is diluted in a 100 ml ofa 0.9% salt solution according to the US Pharmacopoeia under sterileconditions, and the resulting infusion solution is used for infusion.

Example 4 Tablets with Triacetyluridine

[0082] The following ingredients are used for the manufacture of 5000tablets, each of which contains 200 mg of triacetyluridine (for themanufacturing process, see U.S. Pat. No. 5,538,117): Triacetyluridine1000 g Corn starch 680 g Colloidal silica gel 200 g Magnesium stearate20 g Stearic acid 50 g Sodium carboxymethyl starch 250 g Water as muchas is necessary

[0083] A mixture of triacetyluridine, 50 g of cornstarch, and the silicagel is processed together with the starch paste, which consists of 250 gof cornstarch and 2.2 kg of demineralized water, thereby producing amoist mass. This is pressed through a sieve with a mesh size of 3 mm anddried for 30 min. in a fluidized bed dryer at 45° C. The dried granulateis pressed through a sieve with a mesh size of 1 mm, mixed with amixture of 300 g corn starch, the magnesium stearate, the stearic acid,and the sodium carboxymethyl cellulose that was previously passedthrough a 2 mm sieve, and compressed into slightly rounded tablets.

1. The use of active agents that increase the concentration of pyrimidine base building blocks the biosynthesis of nucleic acids in the body for the reduction of side effects of inhibitors of the biosynthesis of nucleic acids or their preliminary stages.
 2. The use according to claim 1 of the active agents for the activation of the biosynthesis of mitochondrial DNA for the reduction of side effects of the inhibitors of the biosynthesis of nucleic acids or their preliminary stages in warm-blooded species, specifically humans.
 3. The use according to claim 1 or 2 of active agents selected from uridine-phosphorylase inhibitors, inhibiting substances of the uridine secretion, compounds that compete with uridine in connection with the renal transport mechanisms, and pyrimidine nucleosides or their prodrugs.
 4. The use according to one of claims 1 to 4, wherein the active agents are selected from among orotic acid, pyrimidine nucleosides and/or prodrugs of pyrimidine nucleosides.
 5. The use according to one of claims 1 to 4, wherein the inhibitors of the biosynthesis of nucleic acids are non-nucleoside-analogous reverse-transcriptase inhibitors.
 6. The use according to one of claims 1 to 4, wherein the active agents are ribo- or deoxyribonucleosides of uracil or cytosine, or respective prodrugs of them.
 7. The use according to claim 6, wherein prodrugs of orotic acid, uracil or cytosine are used as active agents.
 8. The use according to one of claims 6 or 7, wherein the prodrugs are acyl derivates of ribo- or deoxyribonucleosides of uracil or cytosine, preferably those in which one or several of the hydroxy groups in the pentose radical are respectively esterified with an acyl radical, and/or in addition, amino groups, if present, are acylated with acyl.
 9. The use according to claim 8, wherein the prodrugs are those of the following formulas I or II, in which X is hydrogen or —OR₂, and R₁, R₂, R₃ and, if present, R₄ is independently hydrogen or acyl, with at least one of the radicals R₁, R₂, R₃ and R₄ being an acyl:


10. The use according to one of claims 1 to 9, wherein lipodystrophy is treated prophylactically or therapeutically as a side effect.
 11. The use according to one of claims 1 to 9, wherein hyperlacticemia and lactic acidosis and their clinical symptoms are treated prophylactically or therapeutically as a side effect; in particular, acute or chronic hyperlacticemia or lactic acidosis is treated therapeutically.
 12. The use according to one of claims 1 to 9, wherein disorders of spermatogenesis or of the sperm function and/or osteopenia are treated prophylactically or therapeutically as a side effect.
 13. The use according to one of claims 1 to 9, wherein pancreatitis, diminished aerobic capacity and/or disorders of the kidney functions are treated prophylactically or therapeutically as a side effect.
 14. The use according to one of claims 1 to 9, wherein damage to the liver, specifically micro- or macro vesicular steatosis, steato-hepatitis or liver failure, is treated prophylactically or therapeutically as a side effect.
 15. The use according to one of claims 1 to 14 in the context of perinatal transmission prophylaxis with inhibitors of the biosynthesis of nucleic acids or their preliminary stages for the reduction of side effects, specifically on the fetus or the child after birth in the first year of life, especially a newborn.
 16. The use according to one of claims 1 to 9 of active agents for the manufacture of pharmaceutical preparations for the treatment of the side effects named in one of claims 1 or 10 to
 15. 